LOCUS       sequence-270301-        5900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G24100 K4_pECIA14 should be used with bait vector 
            AT5G24100 K4_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5900)
  AUTHORS   Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC, 
            Layeghifard M, Neuhold J, Lehner A, Kong J, Grunwald K, Weinberger 
            N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P, 
            Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir 
            Y
  TITLE     An extracellular network of Arabidopsis leucine-rich repeat receptor
            kinases.
  JOURNAL   Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. 
            Epub 2018 Jan 10.
  PUBMED    29320478
REFERENCE   2  (bases 1 to 5900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature25184"; journalName: "Nature"; date: "2018-01-18- 
            18"; volume: "553"; issue: "7688"; pages: "342-346"
COMMENT     Sequence Label: AT5G24100 K4_pECIA14
FEATURES             Location/Qualifiers
     source          1..5900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        23..449
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     467..520
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1196..1219
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2894..2917
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2933..2950
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2973..2990)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3023..3157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3047..3066)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3101..3120
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3301..3317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3301..3317)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3314..3336)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3325..3341
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3349..3355,3356..3373,3374..3379))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3394..3415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3532..3549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3703..4291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3783..3802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4462..5253,5254..5322))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5085..5104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5323..5427)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5495..5513
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5551..5573)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5673..5692
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5886..5900,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg atgtggtgcc cgatgtgact
       61 agctctttgc tgcaggccgt cctatcctct ggttccgata agagacccag aactccggcc
      121 ccccaccgcc caccgccacc cccatacata tgtggtacgc aagtaagagt gcctgcgcat
      181 gccccatgtg ccccaccaag agctttgcat cccatacaag tccccaaagt ggagaaccga
      241 accaattctt cgcgggcaga acaaaagctt ctgcacacgt ctccactcga atttggagcc
      301 ggccggcgtg tgcaaaagag gtgaatcgaa cgaaagaccc gtgtgtaaag ccgcgtttcc
      361 aaaatgtata aaaccgagag catctggcca atgtgcatca gttgtggtca gcagcaaaat
      421 caagtgaatc atctcagtgc aactaaaggg gggatccgat ctcaatatga agttatgcat
      481 attactggcc gtcgtggcct ttgttggcct ctcgctcggg tctcaggtca ccggtgatct
      541 agccggtgac agacaagccc ttctcgattt cctcaacaac atcattcacc cacggtctct
      601 agcgtggaac acaagcagcc cggtatgcac cacgtggcca ggggtcacat gcgacataga
      661 cggcacacgt gtcactgccc ttcacttgcc cggagcaagt cttctaggag tgatcccacc
      721 tggtactatc agccgtctat cggagcttca gatcctcagt cttcgatcca acggcctacg
      781 agggccgttc ccaattgatt tcttgcagct taagaaactc aaagcaatta gtctcgggaa
      841 caacagattc tctggcccat taccatccga ttacgccacg tggacgaatc tcactgttct
      901 tgatctgtac agtaaccggt ttaacggtag tattcccgcg ggatttgcta atcttaccgg
      961 actcgtctcg cttaacctag ctaaaaactc attctccggt gagattccgg atcttaatct
     1021 ccccggtcta cgcagactta acttttccaa caacaatctc accggatcaa tcccaaattc
     1081 tctgaaaaga ttcgggaact cggctttttc cggcaacaac ttagtcttcg aaaatgcccc
     1141 tcctccggcg gtagtttcat tcaaagagca aaagaaaaac ggcatctata tctctctgga
     1201 ggtgctgttc cagggaccaa gcggcaatcc cggcggcgat ctggcccccc agatgctgcg
     1261 cgagctgcag gagaccaatg ccgccctgca ggacgtgcgc gagctgctgc gccaccgcgt
     1321 gaaggagatc accttcctga agaacaccgt gatggagtgc gacgcctgcg gcatgcagcc
     1381 ctctagaggc tccggctcca tcatcccagt tgaggaggag aacccggact tctggaaccg
     1441 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     1501 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     1561 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     1621 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     1681 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     1741 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     1801 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     1861 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     1921 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     1981 ggacattgac gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     2041 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     2101 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat
     2161 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     2221 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     2281 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     2341 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     2401 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     2461 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     2521 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     2581 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     2641 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     2701 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     2761 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     2821 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggttcta gaggaagcac
     2881 ctccggcgat aacgattaca aggatgatga tgataaggcc agcggcaccg gtcatcatca
     2941 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     3001 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     3061 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3121 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3181 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3241 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     3301 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3361 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3421 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3481 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3541 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4441 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4501 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4561 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4621 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4681 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4741 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4801 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4861 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4921 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4981 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5041 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5101 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5161 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5221 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5281 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5341 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5401 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5461 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5521 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5581 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5641 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5701 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5761 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5821 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5881 gttttcccag tcacgacgtt
//
